immunoserology LECTURE

lesson #7: (week no. 3)

Natural immunity 2. Chemotaxis – cell movement in a

↝ may be: certain direction under the stimulation of
chemical substances
• natural Eg. C5a, C5b, C6, C7
• acquired
Opsonization – coating of antibody and/or
IMMUNITY complement to facilitate phagocytosis
Natural Acquired Eg. C3b, C4b, C5b, Fibronectin,
↝ Physical Barriers ↝ Humoral Immunity Leukotrienes, Immunoglobulin
• B cells, Plasma Engulfment – achieved through amoeboid motion;
Cells, Memory final structure is called vacuole or phagosome
Cells
• Antibodies Degranulation and Digestion
↝ Susceptibility vs. ↝ Cell-Mediated
e. Nonspecific Plasma Proteins
Nonsusceptibility Immunity
Important Terms to Remember:
• T cells, NK,
LAK, K Cells Natural Antibodies
• Lymphokines ↝ Ab present in a host without prior contact with
stimulating agents
↝ Inflammation
↝ Phagocytosis Lysozyme
↝ enzyme found in many types of cells; can also
• Chemotaxis
be found in tears, saliva, and other secretions
vs.
Opsonization
Properdin
↝ Nonspecific ↝ serum protein that exerts bactericidal and
Plasma Proteins viricidal effects in the presence of C3 and Mg

NATURAL/ NONSPECIFIC/ INHERENT/ INNATE Betalysin

IMMUNITY ↝ heat stable cationic substance with
a. Physical Barriers antibacterial activity, found in serum (not in
↝ such as skin and mucous membrane plasma) because it is released by platelets
↝ coughing, sneezing, and vomiting are during coagulation
physiologic responses of nonspecific
immunity
Complement
b. Genetically Controlled Susceptibility and
Nonsusceptibility to Certain Diseases ↝ nonspecific serum proteins that enhance the
Eg. Fy(a-b-) – resistance to malaria effect of antibody; it is activated by Ag-Ab
c. Inflammation interaction
↝ a nonspecific exaggerated physiologic
response which involves a vascular C-Reactive Protein
response and a cellular response by ↝ acute phase plasma protein which increases in
phagocytic cells to infection/injury response to inflammation and tissue necrosis
d. Phagocytosis
↝ engulfment of cells or particulate matter Cytokines
by leukocytes ↝ protein molecules that transmit messages
between the cells
Steps of Phagocytosis
1. Initiation – initiated through tissue INTERFERONS – glycoproteins that has virus-
damage nonspecific antiviral activity

notes by: angelica del pilar

immunoserology LECTURE
lesson #7: (week no. 3)

3 Groups of IFN: IL9 stimulates proliferation

↝ Alpha IFN of T Cell and Mast
– aka Leukocyte IFN Cells
↝ Beta IFN IL10 inhibits cytokine
– aka Epithelial IFN, Fibroblast IFN, synthesis
Fibroepithelial IFN, B-cell
IL11 regulates
Stimulating Factor 2
hematopoiesis
↝ Gamma IFN
– produced by immunologically IL12 NK Cell Stimulating
stimulated lymphocytes, primarily T Factor; enhances
lymphocytes activity of cytotoxic
– a Lymphokine, aka Immune IFN effector T Cells
– main functions:
immunoregulation, enhance NK COMPLEMENT SYSTEM
cells, and activate macrophages ↝ a set of serum that play a role in cytolytic
destruction of cellular antigen by specific antibody
Tumor Necrosis Factor (TNF) is produced ↝ reaction is nonspecific to the target cell
↝ destroyed at 56°C for 30 minutes
mainly by macrophages/monocytes (a
↝ portions of the system contribute to chemotaxis,
monokine).
opsonization, immune adherence, anaphylatoxin
formation and virus neutralization
INTERKULINS – means of communication
between leukocytes
Chemotaxins – C5a, C5b, C6, C7
For Immune Adherence – C3b
IL1 Lymphocyte Activating
Kinin Activator – C2b
Factor
Anaphylatoxins – C3a, C4a, C5a
IL2 T-Cell Growth Factor;
Opsonins – C3b, C4b, C5b
Activates Cytotoxic
Cells, NK Cells, LAK For Virus Neutralization – C4b, C1
Cells
IL3 Multi-colony
THREE PATHWAYS OF ACTIVATION
Stimulating Factor
1. Classical Pathway
(MCSF) Stimulates
↝ requires IgM or IgG for activation; activated by
Hematopoietic Cells
Ag-Ab Complex
IL4 Produced by activated
T cells to promote a. C1 – Recognition Unit
proliferation of B cells ↝ trimolecular complex (C1q, C1r, C1s)
IL5 B Cell Growth Factor held together by Ca++ ions
2; shares functions ↝ C1q attaches to at least two Fc regions
with IL4 (CH2) of Ig adjacent to one another
IL6 IFN beta 2; induces ↝ C1s activates C4
secretion of Ig and
other plasma proteins
IL7 Lymphopoietin 1;
stimulates maturation
of early B and T Cells
IL8 Monocyte-derived
Neutrophil
Chemotactic Factor;
principal inflammatory
cytokine

notes by: angelica del pilar

immunoserology LECTURE
lesson #7: (week no. 3)

b. C4 – First Activation Unit ↝ MBL activates three (3) serine protease:

↝ cleaved into C4a and C4b (C4a –
anaphylatoxin; C4b – attaches to the cell • MASP-1 (unknown function)
membrane) • MASP-2 (cleaves C4 and C2)
↝ C1s interaction with C4 activates C2 • MASP-3 (unknown function)
c. C2 – Second Activation Unit
↝ cleaved into C2a and C2b (note: C2b is ACTIVATING FACTOR: Mannose Groups of
very labile and decays easily with half-life Carbohydrates in Microbial Cell
of 10 minutes if not bound to C4b)
↝ C2a combines with C4b to form C4b2a EFFECTORS: MBP/MBL (Mannan-Binding
complex (C3 convertase) Protein/Lectin), MASP (MBP-Associated Serine
d. C3 – Third Activation Unit/Amplification Protease)
Phase Unit
↝ C3b combines with C4b2a to form
Classical Alternative Lectin
C4b2a3b (C5 convertase)
e. C5, C6, C7, C8, C9 – Membrane Attack Activating Immune LPS Mannose
Phase Units Subs Complexes (bacterial Groups
↝ C8 and C9 are acted on by C5bC6C7 (IgG or capsule) on
complex and insert themselves into the cell IgM) IgA Microbial
membrane resulting in cell lysis Cell

2. Alternative Pathway Of Complement Recognition C1q, C1r, C3, Factor MBP,

Activation Unit C1s B, Factor MASP-1,
a. Factor B – activated into its active form Bb D MASP-2
with the participation of C3; Bb will cleave
C3 into C3a and C3b and will combine with C3 C4b2a C3bBb C4b2a
C3b to form C3bBb Convertase
b. Factor D – cleaves Factor B into Bb in the
presence of C3 and Mg++ ions C5 C4b2a3b C3bBb3b C4b2a3b
Convertase
Therefore,
Factor B + Factor D + C3 + Mg++ → C3bBb
MAC C5b6789
c. Properdin (Factor P) – binds to C3bBb to End Result Cell Lysis
prevent spontaneous decay of the complex
*see pictures/figures on the last page
3. Lectin Pathway (Mbl)
↝ Mannose-Binding Lectin Pathway SYSTEM CONTROL AND REGULATION
↝ Mannan-Binding Lectin Pathway ↝ uncontrolled activation of complement could
↝ provides additional link between innate and cause tissue damage and devastating systematic
acquired immune response effects
↝ non-specific recognition of carbohydrates which ↝ increased complement – little importance
is common constituents of microbial cell walls ↝ decreased complement – more significant
↝ causes:
Mannose-Binding Lectin (MBL) • complement has been abnormally
↝ lectins – proteins that binds carbohydrates consumed
↝ binds to mannose or related sugars to initiate • complement may be decreased or absent
activation of this pathway due to genetic effects
↝ mainly produced by the liver
↝ increase during initial inflammatory C1-Inhibitor (C1INH)
response ↝ dissociates C1r and C1s from C1q
↝ defense of infant – during interval loss of ↝ inhibits activation at first stage of both the
maternal Ab and development of full-fledge Ab classical and lectin pathway
response to specific Ag

notes by: angelica del pilar

immunoserology LECTURE
lesson #7: (week no. 3)

↝ inactivates MASP-2 binding to the MBL-MASP

Complex

Factor I
↝ serine protease that binds to C3b and C4b to
inactivate it

C4 Binding Protein (C4BP)

↝ acts as cofactor with factor I to inactivate C4b

Complement Receptor Type 1 (CR1)

↝ aka CD35 (cofactor with factor I)

Membrane Cofactor Protein (MCP)

↝ aka CD46 (cofactor with factor I)

Decay Accelerating Factor (DAF)

↝ aka CD55 → presence on host cells protects
them from “BYSTANDER LYSIS”

Factor H
↝ competes for Factor B
↝ accelerates dissociation of C3bBb complex on
cell surface
↝ cofactor to factor I

S Protein
↝ vitronectin; prevents attachment of the C5b67 to
cell membrane UNUSUAL NEGATIVE EFFECTS
✓ activate systematically on large scale as in gram
Membrane Inhibitor of Reactive Lysis (MIRL) negative septicemia
↝ aka CD59 ✓ activate by tissue necrosis such as MI
↝ blocks MAC formation ✓ lysis of RBC
↝ binds C8 and prevent C9 to bind to host cell ✓ paroxysmal nocturnal hemoglobinuria
membrane ✓ hereditary angioedema
↝ important in protecting RBC
LABORATORY TESTS
Carboxypeptidase N ↝ Radial Immunodiffusion (RID)
↝ inactivates C3a, C4a, and C5a ↝ Nephelometry
↝ CH50 Assay
COMPLEMENT RECEPTORS
• most commonly used assay for
• CR1 – CD35 measurement of lysis
• CR2 – CD21
• CR3 – CD11b/CD18 ↝ AH50 Assay
• CR4 – CD11c/CD18 • same as CH50 for alternative path
• Collectin Receptors – binds collagen
portion of C1q and generally enhances ↝ Enzyme-Link Immunosorbent Assay (ELISA)
binding of C1q to FC region
• measures classical pathway activation
*see page 101 and 102 of Clinical Immunology and
Serology (A Laboratory Perspective), table 7-3 ↝ CH50 Assay
• most commonly used assay for
measurement of total complement activity

notes by: angelica del pilar

immunoserology LECTURE
lesson #7: (week no. 3)

• amount of complement in serum that can The Membrane Attack Complex (MAC)
cause hemolysis of the 50% reagent Ab-
sensitized SRBC
↝ Complement Fixation Test
• uses antigen as reagent
• test for the presence of complement
proteins
• indicator – Sheep’s Red Blood Cells
• POSITIVE result – (-) hemolysis
• NEGATIVE result – (+) hemolysis

Note:
Professor: Jose Edwardo Mamaat, MD
Prerecorded Lecture: Jose Edwardo Mamaat, MD

notes by: angelica del pilar